GSM6729952: Xyn6h_ii Stationaryl growth; Bacillus subtilis; RNA-Seq
Cell disruption was achieved by bead beating (Precellys 24). After centrifugation, the rough RNA in the upper aqueous phase was ethanol-precipitated, washed twice with 70% ethanol, dried and dissolved in water. DNA was removed by DNAseI (NEB) treatment; the pure total RNA was then extracted by a second-round of P/C/I extraction, following by ethonal-precipitation and 70% ethanol washing, and finally dissolved in water. The total RNA was subjected to MICROBExpress™ Bacterial mRNA Enrichment Kit (Thermo Fisher) to remove >95% of the 16S and 23S rRNA from total RNA. Libraries were constructed using the NEBNext® Ultra™ II Directional RNA Library Prep Kit for Illumina® (New England Biolabs) and NEBNext® Multiplex Oligos for Illumina® (New England Biolabs), according to the manufacturer's protocol.